LABORATORY OF FOOD AND NUTRITIONAL BIOCHEMISTRY AND OF OXIDATIVE STRESS
Belluschi Building
Responsible Prof.ssa Tiziana Bacchetti
Research Activity
The main research activities carried out in this laboratory, focus on the areas reported below:
The research activity is carried out in collaboration with professors and researchers of this Department and of those belonging to other Faculties such as Medicine and Surgery, Agriculture and Engineering (UNIVPM), as well as with researchers from other universities and institutes both national and international.
The results obtained from the different research areas, are reported in numerous publications on international scientific journals and in patents, that can be checked on the staff website.
- Studies on the structure and function of plasma lipoproteins (HDL, LDL) under normal and pathologic conditions
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In this area of research, several methods have been fine tuned to separate the main lipoprotein fractions (very low density lipoproteins, VLDL; low density lipoproteins, LDL and high density lipoproteins, HDL) and their subfractions from human plasma. The study of the composition and physico-chemical properties of lipoproteins (order, polarity) is carried out on lipoproteins isolated from normolipidemic patients, from patients affected by dysmetabolic diseases and in experimental “in vitro” models of ‘pathological’ lipoproteins, such as oxidized, glycosylated, homocysteinylated lipoproteins. Interest in these experimental models, stems from numerous studies showing that lipoproteins, in circulation, are subject to a continuous remodeling of their composition and physic-chemical properties. 
Separation of plasma lipoproteins
- Studies on the antioxidant and antiatherogenic properties of high density lipoproteins (HDL): role of the enzyme paraoxonase
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High density lipoproteins (HDL) play a protective role against the onset of cardiovascular diseases, which has been attributed to their capacity to transport cholesterol from cells to the liver (inverse transport of cholesterol) and to their capacity to inhibit lipid peroxidation of low density lipoproteins (LDL) and of cell membranes. Within this area of research, several methods have been fine tuned to determine the antiatherogenic properties of HDL and to evaluate paraoxonase-1, an antioxidant enzyme present on the surface of HDL. 
Structure of HDL
Antioxidant role of HDL
- Studies on the molecular bases and role of oxidative stress on the pathogenesis of dysmetabolic and neurodegenerative diseases
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Oxidative stress plays a key role in the development and progression of numerous disease states (diabetes, obesity, stroke, multiple sclerosis, psoriasis, celiac disease, neurodegenerative diseases …). In this area of research, several methods have been fine tuned to evaluate the main markers of lipid peroxidation (conjugated dienes, lipid hydroperoxides, ox-LDL) in several biological samples (plasma, liquor, tissues …) both in isolated lipoproteins and in cell cultures. 
Mechanisms involved in promoting lipid peroxidation
Atherogenic role of oxidized LDL
- Studies on the relationship between nutrition and health, and evaluation of the functional properties of food
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Numerous molecules are present in foods, and that play a functional role through different mechanism. In this area, several methods have been fine tuned to evaluate nutritional characteristics of food:
- evaluation of antioxidant levels in different food matrices (total phenols, antioxidant vitamins, carotenoids);
- evaluation of total antioxidant capacity (ABTS assay, ORAC assay, DPPH assay, DMPD assay) of foods and drinks;
- evaluation of markers of oxidative damage in food products;
- evaluation of glycemic index and glycemic load “in vivo” of foods.
This line of research has allowed us to be competent in giving out expert advice to food companies for labeling of their food and nutritional products and to give scientific support for the formulation of different food products with peculiar nutritional characteristics (i.e. low glycemic index, high fibre content, controlled and balanced levels of lipids, carbohydrates, proteins, high levels of antioxidants …). In recent years, we have given technical-scientific support aimed at activating the procedures for approval of health claims by EFSA (European Food Safety Authority) according to the European regulations CE 1924/2006 and CE 353/2008, and to monitor the procedure from the initial steps, to the launch, up to the final phases of approval by the Authority. This has already led to scientific collaborations with different food companies and public institutions (Marche Region, Province of Ancona) for carrying out projects aimed at promoting the nutritional value of foods.
Food Pyramid
Food Label
- Studies of the mechanisms of action of natural and synthetic antioxidants
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IIn this area, the activity of antioxidant molecules present in foods and drinks (polyphenols, phytosterols, carotenoids, antioxidant vitamins) or synthetic antioxidants (eg. nitroxide radicals) is evaluated using different experimental models both in vitro (cell cultures, lipoproteins, biological membranes and model membranes, proteins) and in vivo, both in normal and pathological subjects 
Antioxidant activity of synthetic compounds
Antioxidant Activity of Tea
- Studies on oxidative stress induced by UVA solar radiation
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Solar radiation, especially UVA rays (320-400 nm), are responsible for oxidation of different intra/extracellular biological macromolecules. In this context, the research work carried out is mainly aimed at determining the effects of UVA on human dermal fibroblasts (reached by these rays in the dermis during sun exposure) which induce, photoageing, photocarcinogenesis and immunosuppression. Inhibition of this damage is studied in the presence of both natural and synthetic antioxidants. 
Effetti dei raggi UV sulla pelle
- Studies on the photostability and photoprotective efficacy of commercial UV filters/ sunscreens new UV filters and extracts from plants and marine organisms
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This is one of the main areas of research in the laboratory. Some UV filters present in creams/sunscreens are photounstable and breakdown when exposed to sunlight, generating at times, reactive oxygen species. In this context, the photostability of different commercial UV filters, both organic and inorganic, present in homogeneous systems (solution), in cosmetic formulations and in new vehicles (NLC: Nanostructured Lipid Carriers) exposed to UVA radiation or natural sunlight, are studied. Not only photostability, but also the effects (protective/toxic) of UV filters as well as compounds/extracts from plants and marine organisms, are tested in cosmetic formulations and in biological systems exposed to UV. Cultures of human dermal fibroblasts are used as experimental model, on which several end-points related to UVA-induced oxidative damage are determined. 
Absorbance spectra of sunscreens before and after UVA exposure
Evaluation of the photoprotective efficacy of sunscreens
- Equipment
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• UVA sun lamp (Philips Original Home Solarium, model HB 406/A)
• Heidolph VV2000 Rotavapor with Buchi 461 thermostatted bath
• Centrifuge FALC 4225
• BEL Analytical Balance
• Magnetic Shakers/Hot plates
• pH metre
• BIORAD vertical electrophoresis station
• BIORAD Western Blot station
• Fume hood
• UVIKON 941 Plus Spectrophotometer - Il personale del laboratorio ha inoltre a disposizione le seguenti attrezzature
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• Equipment for cell cultures (Thermo Scientific BB15 Heraeus incubator, Thermo Scientific HERACell 150i incubator with controlled oxygen flux, flow hood, inverted microscope Olympus CK2)
• Spectrophotometer UV/Visible (Shimadzu 2700i) with integrated sphere
• Spectrofluorimeters equipped with polarizers and thermostatted cells (Perkin Elmer LS55, LS50B)
• MyiQ Single Colour Real-Time PCR Detection System (BioRad)
• Microplate Reader (Synergy HT, BioTEK)
• Cytofluorimeter (Millipore Guava easyCyte)
• Ultracentrifuge (Sorvall RC M150GX, Beckman Optima L-90K)
• Refrigerated Centrifugese (Beckman J2-HS, Thermo Scientific Heraeus MEGAFUGE 16R)
• Lyophilizer (Heto DryWinner 6-85)
Staff
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Tiziana Bacchetti (Associate Professor) 071 2204968 |
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Elisabetta Damiani (Associate Professor) 071 2204125 |
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Camilla Morresi (Post-doc) |
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Alessia Luccarini (PhD Student) |
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Luisa Bellachioma (Phd Student) |
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